3895s  (Cell Signaling Technology Inc)

Journal: iScience

Article Title: Context-dependent NMDA receptor dysfunction predicts seizure treatment in mice with human GluN1 variant

doi: 10.1016/j.isci.2025.114301

Figure Lengend Snippet: Potentiating SK channels restores normal NMDAR integration in Grin1 Y647S +/− mice (A) Schematic of potential mechanism showing impaired negative feedback in Y647S +/− neurons due to insufficient Ca 2+ influx compared to WT and the effect of NS309 in boosting SK channel Ca 2+ sensitivity, thereby restoring negative feedback in Y647S +/− neurons. (B) Average NMDAR plateau potential in Y647S +/− neurons at 70 μA with extended tail indicated by red arrow. Application of 10 μM NS309 to the slice restores normal duration and terminates the NMDAR plateau potential in Y647S +/− (Y647S +/− + NS309). (Inset) Restoration of plateau potential duration by NS309 at increasing stimulus intensities in a Y647S +/− neuron. (C) Total width of the NMDAR plateau potential in WT, Y647S +/− , and Y647S +/− + NS309 neurons (∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 10 −4 , Tukey’s post hoc. Black stars: WT vs. Y647S +/− , purple stars: Y647S +/− vs. Y647S +/− + NS309). (D) SK2 channel blocker (Leidab7, 100 nM) prevents NS309 from reducing plateau potential duration in Y647S +/− neurons. (E) Normalized NMDAR plateau width in Y647S +/− neurons with the addition of NS309 and NS309+SK2 blockers (∗ p < 0.05, Sidak’s post hoc).

Article Snippet: NS309 , Tocris , 3895.

Techniques:

Journal: iScience

Article Title: Context-dependent NMDA receptor dysfunction predicts seizure treatment in mice with human GluN1 variant

doi: 10.1016/j.isci.2025.114301

Figure Lengend Snippet: Potentiating SK channels restores normal NMDAR integration in Grin1 Y647S +/− mice (A) Schematic of potential mechanism showing impaired negative feedback in Y647S +/− neurons due to insufficient Ca 2+ influx compared to WT and the effect of NS309 in boosting SK channel Ca 2+ sensitivity, thereby restoring negative feedback in Y647S +/− neurons. (B) Average NMDAR plateau potential in Y647S +/− neurons at 70 μA with extended tail indicated by red arrow. Application of 10 μM NS309 to the slice restores normal duration and terminates the NMDAR plateau potential in Y647S +/− (Y647S +/− + NS309). (Inset) Restoration of plateau potential duration by NS309 at increasing stimulus intensities in a Y647S +/− neuron. (C) Total width of the NMDAR plateau potential in WT, Y647S +/− , and Y647S +/− + NS309 neurons (∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 10 −4 , Tukey’s post hoc. Black stars: WT vs. Y647S +/− , purple stars: Y647S +/− vs. Y647S +/− + NS309). (D) SK2 channel blocker (Leidab7, 100 nM) prevents NS309 from reducing plateau potential duration in Y647S +/− neurons. (E) Normalized NMDAR plateau width in Y647S +/− neurons with the addition of NS309 and NS309+SK2 blockers (∗ p < 0.05, Sidak’s post hoc).

Article Snippet: SK channel positive allosteric modulator NS309 (10 μM, Tocris ) was included in a subset of experiments to restore appropriate duration of dendritic integration in Y647S +/− mice.

Techniques:

Journal: ACS Omega

Article Title: miR-205 Enhances Sensitivity to Genotoxic Agents in HNSCC Cells and Blocks Sphingosine Kinase 2 Action in Tumorigenicity

doi: 10.1021/acsomega.5c06726

Figure Lengend Snippet: miRNA-205 promotes radiosensitivity in SK2-expressing cell lines. (A) Orospheres formed on miR-205 overexpression in NOK-SK2 (pcDNA-miR-205) cells compared to control NOK-SK2 (pcDNA) post-UV exposure. Adhesion sphere assay (B); colony formation was determined by colony formation assay (C); and single spheres (D) on NOK-SK2 (pcDNA-miR-205) cells compared to control NOK-SK2 (pcDNA). (E) Western blot analysis for the protein levels of ZEB2, STAT3, p53Ac, and Snail. Images of spheres were obtained with a microscope. * p < 0.05, ** p < 0.01, *** p < 0.0001.

Article Snippet: Membranes were blocked with 5% nonfat dry milk (BioRad #1706404), incubated with primary antibodies against STAT3 (Cell Signaling #9139, 1:1000), phospho STAT3 (Cell Signaling #9145, 1:2000), NM23-H1 (Santa Cruz #sc-343, 1:1000), ZEB2 (Aviva #ARP39141_P050, 1:1000), p53Ac (Cell Signaling #2525, 1:1000), p53 (Cell Signaling #2527, 1:1000), SNAIL (Cell Signaling #3895, 1:1000), Calnexin (Cell Signaling #2433, 1:1000), and GAPDH (Cell Signaling #2118, 1:1000), overnight at 4 °C.

Techniques: Expressing, Over Expression, Control, Colony Assay, Western Blot, Microscopy